Association between telomere length in the DNA of peripheral blood leukocytes and the propofol dose in anesthesia induction: an observational study

Abstract Introduction: Propofol is a widely used anesthetic and its dose is closely related to aging. Telomere length (TL) is a unique heritable trait, and emerging as a biomarker of aging, health and disease. Telomerase RNA component (TERC) plays an important role in maintaining TL. We proposed a hypothesis that propofol dose in general anesthesia can be predicted by measuring TL before operation, which greatly reduced the risk of anesthesia, especially the elderly. Methods: The association between the propofol dose in anesthesia induction and: TL in the DNA of peripheral blood leukocytes; body weight; sex; difference of the Bispectral Index (BIS) before and after anesthesia induction in patients was evaluated by multivariable linear regression analyses. The mutation at the 5'end or 3'end of TERC was detected. We recruited 100 patients of elective surgery. Results: We found that propofol dose in anesthesia induction was clearly correlated significantly with TL (r = 0.78, p < 0.001), body weight (r = 0.84, p = 0.004), sex (r = 0.83, p= 0.84, p = 0.004), sex (r = 0.83, p = 0.004), and difference of BIS before and after anesthesia induction (r = 0.85, p = 0.029). By comparing the absolute values of standardized regression coefficients (0.58, 0.21, 0.19, and 0.12) of the four variables, it can be seen that TL contributes the most to the propofol dose in anesthesia induction. However, the mutation at the 5' end or 3' end of TERC was not found. Conclusions: These findings provide preliminary evidence that the propofol dose in anesthesia induction was clearly correlated with genetically determined TL. TL may be a promising predictor of the propofol dose, which is beneficial to improve the safety of anesthesia and reduce perioperative complications.

Clinical features of SF3B1 mutation in patients with myelodysplastic syndrome with excess blasts / 中华内科杂志

Objective: To exploring the clinical features of SF3B1-mutated myelodysplastic syndrome with excess blasts (MDS-EB) and analyzing the association between SF3B1 mutation, and efficacy and prognostic significance for patients with MDS-EB. Methods: This was a retrospective case series study. The clinical data of 266 patients with MDS-EB diagnosed in the First Affiliated Hospital of Zhengzhou University between April 2016 and November 2021 were analyzed. The observed indicators included blood routine counts, mutated genes, overall response rate (ORR), overall survival (OS), progression-free survival (PFS), and leukemia-free survival (LFS). The Kaplan-Meier method was used to depict the survival curves. The Log-rank test method was equally used to compare survival across groups and performed the Cox proportional hazard regression model for prognostic analysis. Results: In 266 patients with MDS-EB, 166 (62.4%) were men, and the median age was 57 (17-81) years. Moreover, there were included 26 and 240 patients in the SF3B1-mutated and SF3B1 wild-type groups. Patients in the SF3B1-mutated group were older [median age 65 (51, 69) years vs. 56 (46, 66) years, P=0.033], had higher white blood cell (WBC) counts [3.08 (2.35, 4.78) × 109/L vs. 2.13 (1.40, 3.77) × 109/L], platelet (PLT) counts [122.5 (50.5, 215.0) ×109/L vs. 49.0 (24.3, 100.8) × 109/L], absolute neutrophil counts (ANC) [1.83 (1.01, 2.88) × 109/L vs. 0.80 (0.41, 1.99) × 109/L]and occurrence of DNMT3A mutation [23.1% (6/26) vs. 6.7% (16/240)] (all P<0.05). The ORR were similar in both groups after 2 and 4 cycles of therapy (P=0.348, P=1.000). Moreover, the LFS (P=0.218), PFS (P=0.179) and OS (P=0.188) were similar across the groups. Univariate Cox analysis revealed that SF3B1 mutation did not affect the prognosis of patients with MDS-EB (OS: P=0.193; PFS: P=0.184). Conclusions: Patients with SF3B1 mutation were older, with greater WBC, PLT, and ANC, and SF3B1 mutation easily co-occurred with DNMT3A mutation. From this model, there were no significant differences in efficacy and survival of MDS-EB with or without SF3B1 mutation.

Recent research on platelet-leukocyte aggregates and their role in the pathogenesis of Kawasaki disease / 中国当代儿科杂志

Activated platelets may interact with various types of leukocytes such as monocytes, neutrophils, dendritic cells, and lymphocytes, trigger intercellular signal transduction, and thus lead to thrombosis and synthesis of massive inflammatory mediators. Elevated levels of circulating platelet-leukocyte aggregates have been found in patients with thrombotic or inflammatory diseases. This article reviews the latest research on the formation, function, and detection methods of platelet-leukocyte aggregates and their role in the onset of Kawasaki disease, so as to provide new ideas for studying the pathogenesis of Kawasaki disease.

Asociación entre la variación de glóbulos blancos y las manifestaciones clínicas, anticuerpos e infecciones en adultos con lupus eritematoso sistémico / White blood cells count variation in systemic lupus erythematosus: its association to disease manifestations, antibodies and risk of infections

Introducción: la asociación de leucopenia, linfopenia y neutropenia con la presencia de autoanticuerpos, manifestaciones clínicas e infecciones en pacientes con lupus eritematoso sistémico (LES) no está bien establecida. Los objetivos de este estudio fueron analizar los cambios en los recuentos de leucocitos y linfocitos en pacientes con LES y su asociación con manifestaciones clínicas, autoanticuerpos y riesgo de infecciones. Materiales y métodos: se recolectaron retrospectivamente los valores de leucocitos, linfocitos y neutrófilos. Se agruparon a los pacientes en cinco categorías: recuento de glóbulos blancos normales, leucopenia (persistente o intermitente) y linfopenia (persistente o intermitente). Se registraron las manifestaciones clínicas, los autoanticuerpos acumulados, el daño, la mortalidad, las infecciones y los tratamientos inmunosupresores recibidos de cada paciente. Resultados: se incluyeron 89 pacientes. La linfopenia (89%) fue la anormalidad más frecuente. La leucopenia intermitente y la persistente se detectaron en el 44% y en el 11% de los pacientes, respectivamente. La linfopenia intermitente y la persistente se hallaron en el 44% y en el 45% de los casos. En el análisis univariado, la presencia de rash discoide se asoció a leucopenia (20,4 vs. 5,1; p=0,059) y el tratamiento con mofetil micofenolato a un recuento normal de leucocitos (p=0,046). El compromiso neurológico se asoció a recuento normal de linfocitos (22,2% vs. 0% y 7,5%; p=0,027); los pacientes con anti-RNP (anti ribonucleoproteína nuclear) presentaron más frecuentemente linfopenia persistente (47% vs. 15,4% y 20%; p=0,007). Ninguno de los grupos se asoció a una mayor prevalencia de infecciones. En el análisis multivariado, el mofetil micofenolato se asoció negativamente a leucopenia (OR 0.33 IC 95% 0,1-0,9; p=0,042) y el compromiso neurológico se asoció negativamente a linfopenia (OR 0.08; p=0,022). Conclusiones: en el análisis univariado, el rash discoide se asoció a leucopenia y el anti-RNP a linfopenia. Al ajustar por otras variables significativas, el tratamiento con mofetil micofenolato se asoció a un recuento normal de leucocitos, mientras que las manifestaciones neurológicas se relacionaron a linfocitos normales. No se demostró asociación de las infecciones con ninguno de los grupos.

Introduction: leukopenia, lymphopenia and neutropenia association to clinical manifestations and infections in systemic lupus erythematosus (SLE) is not well defined. The objectives were to analize leucocytes and lymphocytes variations in SLE patients and their association to clinical manifestations, autoantibodies and infections risk. Materials and methods: total white blood cell (WBC) count, lymphocyte, and neutrophils counts were collected retrospectively. Data were grouped into normal WBC cell count, persistent or intermittent leucopenia and lymphopenia. Disease manifestations, accumulated autoantibodies, damage, mortality, infections and immunosuppressants ever received were registered. Results: study sample included 89 patients. Lymphopenia (89%) was the most common abnormality. Intermittent and persistent leukopenia were detected in 44% and 11% cases. Intermittent and persistent lymphopenia were found in 44% and 45% cases. In univariate analysis, discoid rash was associated to leukopenia (20.4 vs 5.1 p=0.059) and mycophenolate treatment to normal leukocyte count (p=0.046). Patients with neurological disorder tended to have normal lymphocyte counts rather than intermittent or persistent lymphopenia (22.2% vs 0% and 7.5% p=0.027); patients with anti-RNP tended to belong to the persistent lymphopenia group (47% vs 15.4% and 20% p=0.007). Infections were not associated to any of the categories. In multivariate analysis mycophenolate was negatively associated to leukopenia (OR 0.33 95% CI 0.1-0.9 p=0.042) while neurological disorder was negatively associated to lymphopenia (OR 0.08 p=0.022). Conclusions: in univariate analysis, discoid rash was associated to leukopenia and anti-RNP to lymphopenia. When adjusted to other significant variables, mycophenolate was related to normal leukocyte while neurological manifestations were to normal lymphocyte counts. Infections were not associated to any of the categories.

LA ADAPTACIÓN DE LA HEMOGLOBINA Y LOS GLOBULOS BLANCOS EN CASOS SEVEROS DE SARS-COV-2 / THE ADAPTATION OF HEMOGLOBIN AND WHITE BLOOD CELLS IN SEVERE CASES OF SARS-COV-2

Dado que el SARS-CoV-2 causa alteraciones en los parámetros hematológicos, muchos estudios se han concentrado en esta área y los han correlacionado con la gravedad de la enfermedad. Se revisó en un estudio retrospectivo de casos graves de COVID-19, en el cual se contabilizaron y documentaron los datos de hemoglobina (Hb) y glóbulos blancos (WBC) de pacientes reclutados en el período de marzo a julio de 2021. Retrospectivamente, los datos extraídos fueron sobre el nivel de Hb y los recuentos de glóbulos blancos de los informes de laboratorio tanto al ingreso como cuando alta de los pacientes. Se calcularon las frecuencias de anemia o eritrocitopenia (Hb <12 g/dL) y leucocitosis (WBC >11×103/µL) en la población estudiada. De 3637 casos de SARS-COV-2, 250 pacientes con condiciones graves de COVID-19 según la definición de la OMS, ingresaron en la UCI o fallecieron, De 250, 80 (32%) ingresaron a UCI, y el resto 170 casos (68%) fallecieron. Mientras que la edad media de los pacientes fue de 55,46±17,49 años, los pacientes con la condición grave tuvieron significativamente mayor edad que aquellos con la condición leve-moderada (edad media de 50,68 frente a 68,59; P: <0,01). El número medio de glóbulos blancos (WBC) fue de 8,88±7,29 x109/L en todos los pacientes con SARS-COV-2, que es significativamente mayor en los casos graves en comparación con aquellos con enfermedad leve-moderada (10,56 frente a 8,95; P: <0,01). El nivel de Hb (<12 g/dL) fue más bajo en el COVID-19 grave que en otros grupos; sin embargo, no fue estadísticamente significativo. El aumento del número de glóbulos blancos y la disminución de la Hb durante la hospitalización de pacientes con SARS-COV-2, puede predecir un mal resultado. Llegamos a la conclusión de que no sólo el número medio de glóbulos blancos era significativamente mayor en los casos severos también la leucocitosis fue un hallazgo común; lo que indica que un mayor número de glóbulos blancos probablemente puede predecir un mal resultado. Asimismo, el nivel de La Hb fue mayor en el grupo leve-moderado; sin embargo, no fue estadísticamente significativo.

SARS-CoV-2 cause alterations in the hematological parameters and many studies have concentrated on this area and correlated these with severity of disease. A retrospective study of severe cases of COVID-19 were reviewed. The data of hemoglobin (Hb) and white blood cells (WBCs) were accounted and documented. Patients recruited at period from March to July, 2021. Retrospectively, data extracted on the Hb level and WBC counts from Lab reports both at admission and when patients discharge. The frequencies of anemia or erythrocytopenia (Hb <12 g/dL) and leukocytosis (WBC >11×103/µL) was calculated in the studied population. Of 3637 SARS-COV-2 cases, 250 patients with severe conditions of COVID-19 as de ne by WHO, which were either admitted to ICU or died. Of 250, 80(32%) were entered to ICU, and the rest 170 cases (68%) who died. While the mean age of the patients was 55.46±17.49 years, patients with the severe condition were signi cantly older than those with the mildmoderate condition (mean age of 50.68 vs. 68.59; P: <0.01). The mean number of white blood cells (WBC) was 8.88±7.29 x109/L in all SARS-COV-2 patients, which is signi cantly higher in the severe cases compared to those with the mild-moderate disease (10.56 vs. 8.95; P: <0.01). The Hb level (<12 g/ dL) was lower in the severe COVID-19 than other groups; however, it was not statistically signi cant. Increased number of WBCs and dropped Hb level during hospitalization of SARS-COV-2 patients may predict a poor outcome. We concluded that not only the mean number of WBCs was signi cantly higher in the severe cases also leukocytosis was a common nding; indicating that an increased number of WBCs may probably predict a poor outcome. Also, the level of Hb was higher in the mild-moderate group; however, it was not statistically signi cant.

Analysis of different platelet-rich fibrin processing / Análise de diferentes processamentos da fibrina rica em plaquetas

Introduction: Platelet-rich fibrin (PRF) is formed by an autologous blood concentrate, with properties that promote cell proliferation and regenerationof bone, gingival and epithelial tissue. Objective: To compare four different procedures for processing as well as obtaining PRF, and analyzing their formation through laboratory techniques. The purpose of the study is to validate a method that produces higher quality PRF for oral surgery use in different branches of dentistry. Material and method: The experiment consisted of collecting blood from 12 volunteers, and processing each patient's sample in 4 different ways. In the following two-stage process analysis, the first, quantitative, step was to analyze the samples' platelet-poor plasma (PPP),with a Neubauer Hemocytometer to count blood components. In the second, qualitative step, the sample's PRF were analyzed under microscopy using histological slides. Result: The statistical analysis of the leukocyte, red blood cell and platelet count did not show any significant distinction when comparing different processes. Histological analysis of the PRF showed fibrin network with platelet aggregation, several leukocytes and presence of red blood cells, with double centrifuged samples presenting more white blood cells than the others. Conclusion: Among the analyzed procedures, the sample centrifuged once at 3000 RPM (1900 G) for 10 min showed the best quality PRF.

Introdução: A fibrina rica em plaquetas (FRP) é formada por um concentrado sanguíneo autólogo, com propriedades de promover a proliferação celular, regeneração de tecido, dentre eles tecido ósseo, gengival e epitelial. Objetivo: Comparar quatro tipos diferentes de processamento e obtenção de FRP, analisando a sua formação através de técnicas laboratoriais. A finalidade do estudo é validar um método que obtenha maior qualidade para o uso em cirurgias nas diversas áreas da odontologia. Material e método: O experimento foi constituído de uma coleta de sangue de 12 voluntários, onde a amostra de cada paciente foi processada de 4 formas. A análise dos processamentos se deu em duas etapas, onde na primeira, quantitativa, foi analisado o plasma pobre em plaquetas (PPP), através de contagem de constituintes sanguíneos em câmara de Neubauer. Na segunda etapa, qualitativa, foram analisadas as FRP em microscopia através de lâminas histológicas. Resultado: A análise estatística da contagem de leucócitos, hemácias e plaquetas não mostraram diferença significativa entre os distintos processamentos. As análises histológicas do FRP mostraram redes de fibrina com agregação plaquetária, inúmeros leucócitos e presença de hemácias, sendo que as amostras de dupla centrifugação apresentaram mais leucócitos que nas demais. Conclusão: Dos procedimentos analisados, a amostra centrifugada a 1 única vez a 3000 RPM (1900 G) por 10 min mostrou-se a melhor qualidade de FRP.

Comparison of initial periodontal therapy and its correlation with white blood cell level in periodontitis patients with or without diabetes mellitus / 北京大学学报(医学版)

OBJECTIVE@#To compare the clinical efficacy of initial periodontal therapy in periodontitis patients with or without type 2 diabetes mellitus and its correlation with white blood cell counts.@*METHODS@#In this study, 32 chronic periodontitis patients without systemic disease (CP group) and 27 chronic periodontitis patients with type 2 diabetes mellitus (CP+DM group) were enrolled. At admission, all the patients went through periodontal examination and fasting blood examination(baseline). Probing depth (PD), attachment loss (AL), bleeding index (BI), plaque index (PLI), white blood cells (WBC) counts and fasting blood glucose (FBG) were recorded respectively, while hemoglobin A1c (HbA1c) was recorded only in CP+DM group. After that, initial periodontal therapy was performed. All the tests were repeated 3 and 6 months after treatment. The changes of periodontal clinical indexes and WBC levels were compared between the two groups before and after treatment, and the correlation between WBC and periodontal clinical indexes and glucose metabolism indexes were analyzed by generalized linear mixed model.@*RESULTS@#At baseline, the periodontal inflammation and destruction were similar in CP and CP+DM group, but the WBC level was significantly higher in CP+DM groups [(6.01±1.26)×109/L vs. (7.14±1.99)×109/L, P=0.01]. After 3 and 6 months of initial periodontal therapy, the mean PD, AL, BI, and PLI in CP+DM and CP groups were significantly lower than the baseline, and the PD in CP+DM group was further decreased by 6 months compared with 3 months [(3.33±0.62) mm vs. (3.61±0.60) mm, P < 0.05]. However, none of these periodontal indexes showed significant difference between the two groups by 3 or 6 months. In CP+DM group, HbA1c at 3 months and 6 months were significantly lower than the baseline [(7.09±0.79)% vs. (7.64±1.16)%, P < 0.05; (7.06±0.78)% vs. (7.64±1.16)%, P < 0.05], and FBG was significantly lower than the baseline by 6 months [(7.35±1.14) mmol/L vs. (8.40±1.43) mmol/L, P < 0.05]. The WBC level in CP group was significantly lower than the baseline level by 3 months [(5.35±1.37)×109/L vs. (6.01±1.26)×109/L, P < 0.05], while that in CP+DM group was significantly lower than the baseline level by 6 months [(6.00±1.37)×109/L vs. (7.14±1.99)×109/L, P < 0.05]. The analysis of genera-lized linear mixed model showed that WBC level was significantly positively correlated with PD and FBG (P < 0.05).@*CONCLUSION@#Initial periodontal therapy can effectively improve the periodontal clinical status of patients with or without type 2 diabetes mellitus, and have benefits on glycemic control in diabetic patients. However, the response of periodontal indexes and WBC level to initial therapy were relatively delayed in diabetic patients. WBC plays an important role in the correlation between diabetes mellitus and periodontitis.

The Expression and Clinical Correlation of the CD68 Factor in Acute Myeloid Leukemia Patients / 中国实验血液学杂志

OBJECTIVE@#To analyze the expression and clinical characteristics of CD68 in bone marrow and peripheral blood of patients with acute myeloid leukemia (AML).@*METHODS@#The expression of CD68 in bone marrow blast cells was detected by four-color flow cytometry in 50 newly diagnosed AML patients and 23 controls. The expression of CD68 in peripheral blood of 85 newly diagnosed AML patients, 29 remission AML patients and 24 controls was detected by ELISA. The correlation between the expression rate of non-M3 AML bone marrow CD68, peripheral blood CD68 concentration and white blood cell count and other clinical data was compared respectively.@*RESULTS@#The median CD68 expression rate in myeloid leukemia cells of non-M3 AML patients was 19.7%, significantly higher than control (0.2%) (P<0.001). The median concentration of non-M3 CD68 in peripheral blood was 67.97 pg/ml, significantly higher than in control (29.94 pg/ml)（P<0.01）. There was no statistically significant difference in the plasma CD68 concentration of the peripheral blood between the newly diagnosed (45.72 pg/ml) and the remission stage (55.12 pg/ml) of non-M3 AML patients by paired analysis (P>0.05). The results showed that the higher the expression rate of CD68 in bone marrow, the higher the count of white blood cells in peripheral blood, and the lower the count of hemoglobin and platelet in peripheral blood. The higher the plasma concentration of CD68 in peripheral blood, the higher the white blood cell count and the lower the complete remission rate.@*CONCLUSION@#The expression of CD68 both in bone marrow and peripheral blood of patients with non-M3 AML is higher than that of control group. Patients with high expression of CD68 show a low rate of complete remission, suggesting that the expression level of CD68 is correlated with treatment response.

White blood segmentation based on dual path and atrous spatial pyramid pooling / 生物医学工程学杂志

The count and recognition of white blood cells in blood smear images play an important role in the diagnosis of blood diseases including leukemia. Traditional manual test results are easily disturbed by many factors. It is necessary to develop an automatic leukocyte analysis system to provide doctors with auxiliary diagnosis, and blood leukocyte segmentation is the basis of automatic analysis. In this paper, we improved the U-Net model and proposed a segmentation algorithm of leukocyte image based on dual path and atrous spatial pyramid pooling. Firstly, the dual path network was introduced into the feature encoder to extract multi-scale leukocyte features, and the atrous spatial pyramid pooling was used to enhance the feature extraction ability of the network. Then the feature decoder composed of convolution and deconvolution was used to restore the segmented target to the original image size to realize the pixel level segmentation of blood leukocytes. Finally, qualitative and quantitative experiments were carried out on three leukocyte data sets to verify the effectiveness of the algorithm. The results showed that compared with other representative algorithms, the proposed blood leukocyte segmentation algorithm had better segmentation results, and the mIoU value could reach more than 0.97. It is hoped that the method could be conducive to the automatic auxiliary diagnosis of blood diseases in the future.

An improved Vision Transformer model for the recognition of blood cells / 生物医学工程学杂志

Leukemia is a common, multiple and dangerous blood disease, whose early diagnosis and treatment are very important. At present, the diagnosis of leukemia heavily relies on morphological examination of blood cell images by pathologists, which is tedious and time-consuming. Meanwhile, the diagnostic results are highly subjective, which may lead to misdiagnosis and missed diagnosis. To address the gap above, we proposed an improved Vision Transformer model for blood cell recognition. First, a faster R-CNN network was used to locate and extract individual blood cell slices from original images. Then, we split the single-cell image into multiple image patches and put them into the encoder layer for feature extraction. Based on the self-attention mechanism of the Transformer, we proposed a sparse attention module which could focus on the discriminative parts of blood cell images and improve the fine-grained feature representation ability of the model. Finally, a contrastive loss function was adopted to further increase the inter-class difference and intra-class consistency of the extracted features. Experimental results showed that the proposed module outperformed the other approaches and significantly improved the accuracy to 91.96% on the Munich single-cell morphological dataset of leukocytes, which is expected to provide a reference for physicians' clinical diagnosis.

Tratamiento de heridas complejas mediante fibrina rica en plaquetas y leucocitos (L-PRF), experiencia inicial de un centro / Treatment of complex wounds using platelet rich fibrin and leukocytes (L-PRF), initial experience of a center

Resumen Introducción El uso de concentrados plaquetarios para el tratamiento de heridas complejas y regeneración tisular está siendo ampliamente utilizado a nivel mundial. Durante el último tiempo, la segunda generación de concentrados plaquetarios, particularmente el L-PRF, ha permitido tratar de manera efectiva a pacientes con esta patología. Debido a su bajo costo y versatilidad, ha sido posible aplicar esta técnica en variadas situaciones clínicas con buenos resultados. El objetivo de este trabajo es presentar nuestra experiencia utilizando L-PRF para la curación de heridas complejas (CHC) como una alternativa al uso de injertos de distinto grado de complejidad. Materiales y Método: Se realizó un análisis prospectivo de una serie de casos de pacientes que fueron sometidos a tratamiento quirúrgico de heridas complejas mediante el uso de L-PRF en el Hospital Santiago Oriente - Luis Tisné Brousse, entre los meses de enero de 2017 y diciembre de 2018. Mediante examen clínico y parámetros de inclusión, de éxito y de fracaso definidos previamente, se evaluó un total de 11 pacientes con heridas complejas a los cuales se les realizó un tratamiento local con injerto de L-PRF. Resultados: _La etiología de las heridas fue variada. 8 (72%) de los casos lograron una epitelización del 100% y 3 (28%) fracasaron. Se identificaron factores predisponentes para el fracaso de la técnica, y también fue posible establecer una relación de predicción de éxito en donde se relaciona una probabilidad alta de epitelización cuando la granulación de la herida ocurre durante los primeros 10 días sobre el injerto de L-PRF. Conclusión: El tratamiento de heridas complejas mediante L-PRF es una alternativa factible, de bajo costo y requerimientos (comparada con el uso de injertos, colgajos y sustitutos dérmicos), es segura en la resolución de heridas complejas, permitiendo disminuir la morbilidad, los costos asociados al tratamiento y estadía hospitalaria.

Introduction: The use of platelet concentrates for the treatment of complex wounds and tissue regenera-tion is being widely used worldwide. During the last time, the second generation of platelet concentrates, particularly L-PRF, has made it possible to effectively treat patients with this pathology. Due to its low cost and versatility, it has been possible to apply this technique in various clinical situations with good results. The objective of this work is to present our experience using L-PRF for the healing of complex wounds (HCC) as an alternative to the use of grafts of different degrees of complexity. Materials and Method: A prospective analysis was carried out with a series of cases who underwent surgical treatment of complex wounds using L-PRF at Santiago Oriente - Luis Tisné Brousse Hospital, between the months of January 2017 and December 2018. Through clinical examination and previously defined inclusion, success, and failure parameters, a total of 11 patients with complex wounds were evaluated who underwent local treatment with an L-PRF graft. Results: The etiology of the wounds was varied. 8 (72%) of the cases achieved 100% epithelialization and 3 (28%) failed. Predisposing factors for the failure of the technique were identified, and it was also possible to establish a predictive relationship of success where a high probability of epithelialization is related when the granulation of the wound occurs during the first 10 days on the L-PRF graft. Conclusion: The treatment of complex wounds using L-PRF is a feasible alternative, with low cost and requirements (compared to the use of grafts, flaps and dermal substitutes) and safe in the resolution of complex wounds, allowing to reduce morbidity, the costs associated with treatment and hospital stay.

Leukocyte reduction filters as an alternative source of peripheral blood leukocytes for research

ABSTRACT Introduction: Peripheral blood leukocytes are a suitable cell model for science research. However, blood samples from healthy volunteers are limited in volume and difficult to obtain due to the complexity of volunteer recruitment. Objective: Therefore, it is urgent to find an alternative source of peripheral blood leukocytes. Method: One of the possibilities is the use of leukocyte reduction filters (LRFs) in blood banks that is used for preparation of leukoreduced blood products. More than 90% of the leukocytes are trapped in the leukofilters allowing the desired blood product to pass through. Results: It has been reported that the biological function of leukocytes collected from the filters are no different from those isolated from buffy coats, leukapheresis products and whole blood (WB) cells. Moreover, LRFs are waste products that are discarded after leukoreduction. Conclusion: Thus, leukofilters represent an economic source of human cell populations that can be used for a variety of investigative purposes, with no cost. In the present study, we reviewed the different usage of LRFs in the research, clinical and commercial applications.

Relação entre exercício físico e sistema imunológico / Relationship between exercise and the immune system

O corpo humano tende sempre a procurar um estado de homeostase, buscando o equilíbrio entre todos os sistemas. O exercício físico está presente na rotina diária de indivíduos, mesmo com objetivos diferentes, porém a influência no sistema imunológico não é muitas vezes abordada como fator relevante. O sistema imune é responsável por proteger o organismo contra infecções e doenças, podendo ser modulado perante a resposta de exercícios físicos regulares. Tendo em vista que, atualmente, existe uma preocupação maior em tornar e manter a imunidade eficiente, a prática regular e moderada do exercício pode contribuir para uma maior eficácia desse sistema, dessa forma, podendo ser considerada uma proteção ao corpo humano. O objetivo dessa revisão foi sintetizar os dados de estudos presentes na literatura que demonstram a influência do exercício físico na resposta do sistema imunológico, tornando possível compreender as alterações moleculares, fisiológicas, metabólicas e celulares que levam a um tipo específico de resposta do organismo humano.

The human body always tends to seek a homeostasis state, trying to balance all systems. Physical exercise is present in the routine of individuals even with different goals, but the influence in the immune system isnt a relevant factor. The immune system is responsible for protecting the human body against some infections and diseases, and could be modulated in response by some regular physical exercise. At the moment there is a greater concern to keep efficient immunity, a practice of regular and moderate exercise can contribute to a better effectiveness of this system, thus, it can be considered a form of protection to the human body. The objective of this review was to synthesize some data from any studies presented in the literature that demonstrate the influence of physical exercise on the immune system response. Making it possible to understand the molecular mechanisms, physiological, metabolic and cellular changes that turn to a specific type of response in the human body.

Effects of elderflower extract enriched with polyphenols on antioxidant defense of salmon leukocytes

BACKGROUND: In fish farming, the plant extracts containing antioxidant compounds have been added to the diet for enhancing pathogen resistance. In vitro studies evaluating the antioxidant effect of herbal extracts on fish cell models have focused on ROS production and the respiratory burst mechanism. However, the effects on enzymatic antioxidant defense on salmon leukocytes have not been evaluated. This study aims to evaluate the enzymatic antioxidant defense and ROS-induced cell damage in Salmon Head Kidney-1 (SHK-1) cell line exposed to polyphenol-enriched extract from Sambucus nigra flowers. RESULTS: Firstly, the Total Reactive Antioxidant Power (TRAP) assay of elderflower polyphenol (EP) was evaluated, showing 459 and 489 times more active than gallic acid and butyl hydroxy toluene (BHT), respectively. The toxic effect of EP on salmon cells was not significant at concentrations below 120 mg/ mL and no hemolysis activity was observed between 20 and 400 mg/mL. The treatment of SHK-1 cell line with EP decreased both the lipid peroxidation and protein oxidation induced by H2O2, which could be associated with decreasing oxidative stress in the SHK-1 cells since the GSH/GSSG ratio increased when only EP was added. CONCLUSIONS: These results suggest that plant extracts enriched with polyphenols could improve the enzymatic antioxidant defense of salmon leukocytes and protect the cells against ROS-induced cell damage

Effect of Astaxanthin on Antioxidant Enzyme Activities in Suspended Leukocyte-Depleted Red Blood Cells Stored for Transfusion / 中国实验血液学杂志

OBJECTIVE@#To observe the effect of astaxanthin (ASTA) on the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in suspended leukocyte-depleted red blood cells stored for transfusion.@*METHODS@#The suspended leukocyte-depleted red blood cells were randomly divided into group A, B, C and D. The ASTA was added into preservation solution of suspended leukocyte-depleted red blood cells of group B, C and D with the final concentration 5, 10 and 20 μmol/L, respectively, while DMSO was added into cells of group A in the same volume. After 7, 14, 28 and 42 days of storage, the reactive oxygen species (ROS) content in red blood cells was detected by fluorescence microplate reader, malondialdehyde (MDA) content was detected by thiobarbituric acid (TBA) method, activity of SOD was detected by xanthine oxidase method, the activity of CAT was detected by visible light method, and activity of GSH-Px was detected by colorimetry.@*RESULTS@#After 7, 14, 28 and 42 days of storage, the contents of ROS and MDA in suspended red blood cells of group B, C and D were significantly lower(P<0.05), while the activities of SOD and GSH-Px were higher than those of group A(P<0.05); and CAT activity in cells treated by ASTA was significantly higher at 28 and 42 days of storage in comparison with that of group A(P<0.05). There were positive correlations between the ROS, MDA content in suspended red blood cells of group A, B, C, D and storage time(P<0.01), while negative correlation between SOD, CAT, GSH-Px activity and storage time(P<0.01).@*CONCLUSION@#ASTA can decrease the oxidative stress level and peroxide damage degree by increasing the antioxidant enzyme activities in suspended leukocyte-depleted red blood cells during storage.

Cytometric blood phenotyping in free-living white-eyed parakeet (Psittacara leucophthalmus) / Fenotipagem citométrica do sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus)

This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.(AU)

Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.(AU)

Relationship between Leukocytes Derived Microparticles and Minimal Residual Disease and Prognosis of Acute Myeloid Leukemia / 中国实验血液学杂志

OBJECTIVE@#To detect the relationship between leukocytes derived microparticle (CD45@*METHODS@#The expression of CD45@*RESULTS@#The percentages of CD45@*CONCLUSION@#High level of CD45

MicroRNA-135a in ABCA1-labeled Exosome is a Serum Biomarker Candidate for Alzheimer's Disease / 生物医学与环境科学（英文）

Objective@#In the present study, the ABCA1 was used as a label to capture specific exosomes, the level of ABCA1-labeled exosomal microRNA-135a (miR-135a) was evaluated for the diagnosis of Alzheimer's disease (AD), especially in patients with early stages of AD.@*Methods@#This is a preliminary research focused on the levels of ABCA1 in WBCs, RBCs, HT-22 cells, and neuron cells. The diagnostic value of ABCA1-labeled exosomal miR-135a was examined using the CSF and serum of APP/PS1 double transgenic mice, and 152 patients with SCD, 131 patients with MCI, 198 patients with DAT, and 30 control subjects.@*Results@#The level of ABCA1 exosomes harvested from HT-22 cells and neuron culture medium was significantly higher compared to that of RBCs and WBCs ( @*Conclusion@#This study outlines a method to capture specific exosomes and detect them using immunological methods, which is more efficient for early diagnosis of AD.

O fator de virulência relacionado ao H. pylori influencia a expressão de quimiocinas CXCL10, CCL17, CCL20, CCL22, e seus receptores por células mononucleares de sangue periféricos de pacientes com úlceras pépticas / The h. pylori-related virulence factor caga influences the expression of chemokines cxcl10, ccl17, ccl20, ccl22, and their receptors by peripheral blood mononuclear cells from peptic ulcer patients

ABSTRACT BACKGROUND: During the Helicobacter pylori (HP) infection, the infiltration of the leukocytes into stomach mucosa is directed by locally produced chemokines that play a decisive role in infection outcome. The CagA is the most potent virulence factor of HP, so that the infection with CagA + strains is associated with more severe complications than infection with CagA - HP. OBJECTIVE: The aim was to determine the expression of chemokines CXCL10, CCL17, CCL20 and CCL22, and their receptors by CagA + HP- and CagA - HP-derived crude extract (HP-CE)-stimulated peripheral blood mononuclear cells (PBMCs) from peptic ulcer (PU) patients. METHODS: The serum and the PBMCs were collected from 20 HP-infected PU patients, 20 HP-infected asymptomatic subjects (HIA) and 20 non-infected healthy subjects (NHS). The PBMCs were cultured in absence of stimulator or with 10 µg CagA + HP crude extract (CagA + CE), 10 µg CagA - HP crude extract (CagA - CE). Chemokines and receptors were measured by ELISA and real time-PCR respectively. RESULTS: In PU patients, the production of chemokines CXCL10, CCL17, CCL20 and CCL22, and the expression of chemokine receptors CXCR3, CCR4 and CCR6 by CagA + CE-induced PBMCs were significantly higher than non-stimulated and CagA - CE stimulated cultures. The CXCL10 production by CagA + CE stimulated PBMCs from HIA subjects was significantly higher than the equal cultures from PU and NHS groups. The CCL17 and the CCL20 production by non-stimulated, CagA + CE stimulated, and CagA - CE stimulated PBMCs from PU subjects were significantly higher than the equal cultures from NHS and HIA groups. The CCL22 production by non-stimulated, CagA + CE stimulated and CagA - CE stimulated PBMCs from NHS group were significantly higher than the equal cultures from HIA and PU groups. The CagA + CE stimulated PBMCs from HIA subjects expressed lower amounts of CCR6 in comparison with CagA + CE stimulated PBMCs from NHS and PU groups. The serum levels CXCL10 and CCL20 in PU and HIA groups were significantly higher than NHS subjects. NHS and HIA groups displayed higher serum levels of CCL22 in comparison with PU patients. CONCLUSION: Results indicated that the CagA status of bacterium influence the expression of chemokines and receptors by HP-CE stimulated PBMCs from PU patients.

RESUMO CONTEXTO: Durante a infecção por Helicobacter pylori (HP), a infiltração dos leucócitos na mucosa estomacal é dirigida por quimiocinas produzidas localmente que desempenham um papel decisivo no resultado da infecção. O CagA é o fator de virulência mais potente do HP, de modo que a infecção com cepas CagA + está associada a complicações mais graves do que a infecção com CagA - HP. OBJETIVO: O objetivo foi determinar a expressão das quimiocinas CXCL10, CCL17, CCL20 e CCL22, e seus receptores por CagA + HP- e CagA - extrato bruto (EB) derivado de HP (HP-EB) de células mononucleares do sangue periférico (CMSP) de pacientes com úlcera péptica (UP). MÉTODOS: O soro e as CMSP foram coletados de 20 pacientes com UP infectados pelo HP, 20 indivíduos assintomáticos infectados pelo HP (AI-HP) e 20 indivíduos saudáveis não infectados pelo HP (NI-HP). As CMSP foram cultivadas na ausência de estimulador ou com extrato bruto CagA + HP de 10 μg (CagA + EB), 10 μg CagA - extrato bruto HP (CagA - EB). Quimiocinas e receptores foram medidos por ELISA e PCR em tempo real, respectivamente. RESULTADOS: Em pacientes com UP a produção de quimiocinas CXCL10, CCL17, CCL20 e CCL22, e a expressão dos receptores de quimiocina CXCR3, CCR4 e CCR6 por CagA + CMSP induzidos pelo EB foram significativamente maiores do que as culturas não estimuladas e CagA - EB estimulados. A produção de CXCL10 por CagA + EB estimulou as CMSP de sujeitos AI-HP em proporção significativamente maior do que as culturas iguais dos grupos UP e NI-HP. A produção de CCL17 e CCL20 por grupos não estimulados, CagA + EB estimulado, e CagA - EB estimulou CMSP de sujeitos com UP e foram significativamente superiores às culturas iguais dos grupos NI-HP e AI-HP. A produção de CCL22 por grupos não estimulados, CagA + EB estimulado e CagA - EB estimulado pelo grupo NI-HP foram significativamente maiores do que as culturas iguais dos grupos AI-HP e PU. O CagA + EB estimulou as CMSP dos sujeitos do AI-HP, expressando menores quantidades de CCR6 em comparação com as CMSP estimuladas pelo CagA + EB de grupos NI-HP e UP. Os níveis sanguíneos de CXCL10 e CCL20 nos grupos UP e AI-HP foram significativamente superiores aos dos sujeitos do NI-HP. Os grupos NI-HP e AI-HP apresentaram níveis sanguíneos mais elevados de CCL22 em comparação com pacientes com UP. CONCLUSÃO: Os resultados indicaram que o estado CagA da bactéria influencia a expressão de quimiocinas e receptores por HP-EB estimulados CMSP de pacientes com UP.